Radiomitigating pharmaceutical formulations

ABSTRACT

The present disclosure relates to compounds of Formula (I) and (II), compositions containing the compounds (alone or in combination with other agents), and their use to prevent, mitigate or treat a) damage induced by ionizing radiation, b) inflammation or c) cancer.

RELATED APPLICATIONS

This application claims the benefit of priority to U.S. ProvisionalPatent Application No. 61/820,447, filed May 7, 2013, which applicationis hereby incorporated by reference in its entirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with Government support under AI067769, awardedby the National Institutes of Health. The Government has certain rightsin the invention.

BACKGROUND Radiomitigation

The tragic nuclear power plant accidents in Fukushima, Japan causedsevere leaks of radioactive Iodine-131 and Cesium-137 and a subsequentwidespread exposure scare of radiation. In addition, the global use andstorage of radioactivity is increasing rapidly. Millions of radioactivesealed sources are used around the world for legitimate and beneficialcommercial applications such as cancer treatment, food and bloodsterilization, oil exploration, remote electricity generation,radiography, and scientific research. These applications use isotopessuch as Cesium-137, Cobalt-60, Strontium-90, Americium-241, Iridium-192,Plutonium-238, Plutonium-239, Curium-244, Radium-226, andCalifornium-252. Many of these radiological sources at sites around theworld are no longer needed and have been abandoned or orphaned; othersare poorly guarded, making the risk of theft or sabotage significant.Currently, there are tens of thousands of civilian locations worldwidewith radioactive material, about 5,000 of which contain sources of 1,000curies or greater (Office of Global Threat Reduction (NA-21), GTRIStrategic Plan, release date January 2007. 955 L'Enfant Plaza,Washington, D.C. 20585. Iliopulos, Ioanna et al. The Office of GlobalThreat Reduction: reducing the global threat from radiological dispersaldevices. 2007. JNMM Volume 35 Issue 3 PP 36-40). Beyond the publicsafety concerns are the clinical implications of radiation use.

Outside the radiation therapy clinic there is also significant relevanceto identifying and characterizing novel compounds that protect ceilsfrom radiation induced cell death.

Fundamental to radiation exposure and injury is DNA strand breaks,resulting in genetic instability and DNA deletions which are involved incell death, cellular dysfunction, as well as long-term consequences suchas birth defects and cancer.

Discovery of compounds that are capable of mitigating the process ofnormal tissue damage from radiation during radiotherapy, accidents, orterrorist attacks is of importance. Most currently available treatmentsfor radiation exposure are free radical scavengers that reduce initialradiation-induced DNA damage and work best if added just before or atthe time of irradiation. Because of this, these compounds are notpractical countermeasures in a radiation incident. In that case, thesearch for radiomitigators—agents with robust, prolonged efficacy, broadspecificity, and minimal toxicity that could protect a large populationin the event of a radiological emergency is of importance.

SUMMARY OF THE INVENTION

The present invention provides compounds having structures as disclosedherein. Preferred compounds are radiomitigating, and thus can be used inthe prevention, mitigation and treatment of radiation injury, and othermedical conditions related to exposure to ionizing radiation.

The subject compounds are also useful for treating or preventinginflammatory disease and for treating or preventing cancer or otherhyperproliferative conditions.

In one aspect, the invention provides compounds represented by generalformula I or a pharmaceutically acceptable salt, ester, or prodrugthereof:

wherein:

A⁵ is a secondary or tertiary amine (i.e., thereby forming asulfonamide), and

A⁶ is a substituted or unsubstituted and or heteroaryl group, preferablywherein the aryl or heteroaryl group bears at least one substituentincluding a nitro substitutent, e.g., disposed at a position distal tothe sulfonyl.

In certain embodiments, A⁵ is a heterocyclic amine, such as apiperidine, piperazine, or morpholine ring, while in other embodiments,the amine is acyclic and/or the nitrogen atom bound to the sulfonyl isnot included in any ring that may be present in A⁵.

In certain embodiments, the invention provides compounds represented bygeneral formula II or a pharmaceutically acceptable salt, ester orprodrug thereof:

wherein:

X is N or —C(H)— preferably N;

Y¹ and Y² are each independently lower alkyl or Y¹ and Y² taken togetherwith X form a heterocyclyl ring system, such as

wherein

X is N;

G is selected from N or —C(H)—, preferably N;

Z is absent or selected from substituted or unsubstituted alkyl,heteroalkyl, alkenyl, or alkynyl; and

R⁴ is hydrogen or selected from substituted or unsubstituted aryl (e.g.,phenyl) and heteroaryl, and

R⁵ and R⁶ are each independently absent or lower alkyl.

In other embodiments, X, Y¹ and Y² taken together form a ring system

wherein X is —C(H)—, and

R⁴ is selected from substituted or unsubstituted aryl (e.g., phenyl) andheteroaryl, such as a halogen-substituted phenyl group, e.g.,4-fluorophenyl or 3-chlorophenyl.

In certain embodiments, Y¹ and Y² are each ethyl.

In certain preferred embodiments, Y¹ and Y² taken together form apiperazine ring.

In certain preferred embodiments, Z is absent.

In certain embodiments, the compound of Formula II has a structure ofone of compounds 1-7. However, in certain preferred embodiments of thecompounds, compositions, uses, and methods disclosed herein, compounds1-12 (or even compounds 1-12 and compounds P1, P2, and P3) are excluded.

In certain embodiments, compounds of the invention may be prodrugs ofthe compounds of formula I or II, e.g., wherein a hydroxyl in the parentcompound is presented as an ester or a carbonate, or carboxylic acidpresent in the parent compound is presented as an ester. In certain suchembodiments, the prodrug is metabolized to the active parent compound invivo (e.g., the ester is hydrolyzed to the corresponding hydroxyl, orcarboxylic acid).

In certain embodiments, compounds of the invention may be racemic. Incertain embodiments, compounds of the invention may be enriched in oneenantiomer. For example, a compound of the invention may have greaterthan 30% ee, 40% ee, 50% ee, 60% ee,70% ee, 80% ee, 90% ee, or even 95%or greater ee. In certain embodiments, compounds of the invention mayhave more than one stereocenter. In certain such embodiments, compoundsof the invention may be enriched in one or more diastereomer. Forexample, a compound of the invention may have greater than 30% de, 40%de, 50% de, 60% de, 70% de, 80% de, 90% de, or even 95% or greater de.

In certain embodiments, the present invention relates to methods oftreatment with a compound of formula I or II, or a pharmaceuticallyacceptable salt thereof. In certain embodiments, the therapeuticpreparation may be enriched to provide predominantly one enantiomer of acompound (e.g., of formula I or II). An enantiomerically enrichedmixture may comprise, for example, at least 60 mol percent of oneenantiomer, or more preferably at least 75, 90, 95, or even 99 molpercent. In certain embodiments, the compound enriched in one enantiomeris substantially free of the other enantiomer, wherein substantiallyfree means that the substance in question makes up less than 10%, orless than 5%, or less than 4%, or less than 3%, or less than 2%, or lessthan 1% as compared to the amount of the other enantiomer, e.g., in thecomposition or compound mixture. For example, if a composition orcompound mixture contains 98 grams of a first enantiomer and 2 grams ofa second enantiomer, it would be said to contain 98 mol percent of thefirst enantiomer and only 2% of the second enantiomer.

In certain embodiments, the therapeutic preparation may be enriched toprovide predominantly one diastereomer of a compound (e.g., of formula Ior II). A diastereomerically enriched mixture may comprise, for example,at least 60 mol percent of one diastereomer, or more preferably at least75, 90, 95, or even 99 mol percent.

In certain embodiments, the present invention relates to methods oftreatment with a compound of formula I or II, or a pharmaceuticallyacceptable salt thereof. In certain embodiments, the therapeuticpreparation may be enriched to provide predominantly one enantiomer of acompound (e.g., of formula I or II). An enantiomerically enrichedmixture may comprise, for example, at least 60 mol percent of oneenantiomer, or more preferably at least 75, 90, 95, or even 99 molpercent. In certain embodiments, the compound enriched in one enantiomeris substantially free of the other enantiomer, wherein substantiallyfree means that the substance in question makes up less than 10%, orless than 5%, or less than 4%, or less than 3%, or less than 2%, or lessthan 1% as compared to the amount of the other enantiomer, e.g., in thecomposition or compound mixture. For example, if a composition orcompound mixture contains 98 grams of a first enantiomer and 2 grams ofa second enantiomer, it would be said to contain 98 mol percent of thefirst enantiomer and only 2% of the second enantiomer.

In certain embodiments, the therapeutic preparation may be enriched toprovide predominantly one diastereomer of a compound (e.g., of formula Ior II). A diastereomerically enriched mixture may comprise, for example,at least 60 mol percent of one diastereomer, or more preferably at least75, 90, 95, or even 99 mol percent.

In certain embodiments, the present invention provides a pharmaceuticalpreparation suitable for use in a human patient, comprising any of thecompounds shown above (e.g., a compound of the invention, such as acompound of formula I or II), and one or more pharmaceuticallyacceptable excipients. In certain embodiments, the pharmaceuticalpreparations may be for use in treating or preventing a condition ordisease as described herein. In certain embodiments, the pharmaceuticalpreparations have a low enough pyrogen activity to be suitable for usein a human patient.

Compounds of any of the above structures may be used in the manufactureof medicaments for the treatment of any diseases or conditions disclosedherein.

In certain embodiments, the compounds of Formula I or II mitigate tissuedamage induced by exposure to ionizing radiation and/or inhibitinflammation.

In one aspect, the invention provides a pharmaceutical compositioncomprising a compound as disclosed herein and a pharmaceuticallyacceptable excipient or solvent. In certain embodiments, apharmaceutical composition may comprise a prodrug of a compound asdisclosed herein.

In another aspect, the invention provides a method of mitigating theeffect of ionizing radiation on a cell, organ, tissue, or organism bycontacting the cell, organ, tissue, or organism with at least onecompound shown in Table 1. The cell, organ, tissue, or organism may becontacted with a compound shown in Table 1 before, during, or afterexposure to ionizing radiation.

In some embodiments, the compound may be administered prophylactically,i.e., before exposure to ionizing radiation, for example, prior tocancer radiation therapy or X-ray. In some embodiments, the compound maybe administered during exposure, or upon repeated exposure to ionizingradiation. In some embodiments, the compound may be administered afterexposure to ionizing radiation, or after the initiation of exposure toradiation.

When administering a compound of Formula I or II to an organism, thecompound may be administered by any suitable means. In some embodiments,the compounds or formulations are administered orally. In someembodiments, the compounds or formulations are administered byinjection, e.g. subcutaneous, parenteral, or intravenous, injections. Insome embodiments, the compound may be administered in combination withother potential mitigators. In certain embodiments, compounds of FormulaI or II are administered in conjunction with other therapies, such asradiation therapy, other anti-inflammatory compounds, or otheranticancer drugs.

In certain embodiments, a method of the invention may comprisecontacting a cell with a prodrug of a compound as disclosed herein.

Definitions

The terms “a,” “an,” “the” and similar referents used in the context ofdescribing the present invention are to be construed to cover both thesingular and the plural, unless otherwise indicated herein or clearlycontradicted by context. All methods described herein can be performedin any suitable order unless otherwise indicated herein or otherwiseclearly contradicted by context. The use of any and all examples, orexemplary language (e.g., “such as”) provided herein is intended merelyto better illuminate the present invention and does not pose alimitation on the scope of the invention otherwise claimed. No languagein the present specification should be construed as indicating anyunclaimed element is essential to the practice of the invention.

The term “acyl” is art-recognized and refers to a group represented bythe general formula hydrocarbylC(O)—, preferably alkylC(O)—.

The term “acylamino” is art-recognized and refers to an amino groupsubstituted with an acyl group and may be represented, for example, bythe formula hydrocarbylC(O)NH—.

The term “acyloxy” is art-recognized and refers to a group representedby the general formula hydrocarbylC(O)O—, preferably alkylC(O)O—.

The term “alkoxy” refers to an alkyl group, preferably a lower alkylgroup, having an oxygen attached thereto. Representative alkoxy groupsinclude methoxy, ethoxy, propoxy, tert-butoxy and the like.

The term “alkoxyalkyl” refers to an alkyl group substituted with analkoxy group and may be represented by the general formulaalkyl-O-alkyl.

The term “alkenyl”, as used herein, refers to an aliphatic groupcontaining at least one double bond and is intended to include both“unsubstituted alkenyls” and “substituted alkenyls”, the latter of whichrefers to alkenyl moieties having substituents replacing a hydrogen onone or more carbons of the alkenyl group. Such substituents may occur onone or more carbons that are included or not included in one or moredouble bonds. Moreover, such substituents include all those contemplatedfor alkyl groups, as discussed below, except where stability isprohibitive. For example, substitution of alkenyl groups by one or morealkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups iscontemplated.

An “alkyl” group or “alkane” is a straight chained or branchednon-aromatic hydrocarbon which is completely saturated. Typically, astraight chained or branched alkyl group has from 1 to about 20 carbonatoms, preferably from 1 to about 10 unless otherwise defined. Examplesof straight chained and branched alkyl groups include methyl, ethyl,n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl,pentyl and octyl. A C₁-C₆ straight chained or branched alkyl group isalso referred to as a “lower alkyl” group. An alkyl group with two openvalences is sometimes referred to as an alkylene group, such asmethylene, ethylene, propylene and the like.

Moreover, the term “alkyl” (or “lower alkyl”) as used throughout thespecification, examples, and claims is intended to include both“unsubstituted alkyls” and “substituted alkyls”, the latter of whichrefers to alkyl moieties having substituents replacing a hydrogen on oneor more carbons of the hydrocarbon backbone. Such substituents, if nototherwise specified, can include, for example, a halogen, a hydroxyl, acarbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl),a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate),an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, anamino, an amido, an amidine, an imine, a cyano, a nitro, an azido, asulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, asulfonamide, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic orheteroaromatic moiety. It will be understood by those skilled in the artthat the moieties substituted on the hydrocarbon chain can themselves besubstituted, if appropriate. For instance, the substituents of asubstituted alkyl may include substituted and unsubstituted forms ofamino, azido, imino, amido, phosphoryl (including phosphonate andphosphinate), sulfonyl (including sulfate, sulfonamide, sulfamoyl andsulfonate), and silyl groups, as well as ethers, alkylthios, carbonyls(including ketones, aldehydes, carboxylates, and esters), —CF₃, —CN andthe like. Exemplary substituted alkyls are described below. Cycloalkylscan be further substituted with alkyls, alkenyls, alkoxys, alkylthios,aminoalkyls, carbonyl-substituted alkyls, —CF₃, —CN, and the like.

The term “C_(x-y)” when used in conjunction with a chemical moiety, suchas, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant toinclude groups that contain from x to y carbons in the chain. Forexample, the term “C_(x-y)alkyl” refers to substituted or unsubstitutedsaturated hydrocarbon groups, including straight-chain alkyl andbranched-chain alkyl groups that contain from x to y carbons in thechain, including haloalkyl groups such as trifluoromethyl and2,2,2-tirfluoroethyl, etc. C₀ alkyl indicates a hydrogen where the groupis in a terminal position, a bond if internal. The terms“C_(2-yl)alkenyl” and “C_(2-y)-alkynyl” refer to substituted orunsubstituted unsaturated aliphatic groups analogous in length andpossible substitution to the alkyls described above, but that contain atleast one double or triple bond respectively. As applied toheteroalkyls, “C_(x-y)” indicates that the group contains from x to ycarbons and heteroatoms in the chain. As applied to carbocyclicstructures, such as aryl and cycloalkyl groups, “C_(x-y)” indicates thatthe ring comprises x to y carbon atoms. As applied to heterocyclicstructures, such as heteroaryl and heterocyclyl groups, “C_(x-y)”indicates that the ring contains from x to y carbons and heteroatoms. Asapplied to groups, such as aralkyl and heterocyclylalkyl groups, thathave both ring and chain components, “C_(x-y)” indicates that the ringand the chain together contain from x to y carbon atoms and, asappropriate heteroatoms.

The term “alkylamino”, as used herein, refers to an amino groupsubstituted with at least one alkyl group.

The term “alkylthio”, as used herein, refers to a thiol groupsubstituted with an alkyl group and may be represented by the generalformula alkylS-.

The term “alkynyl”, as used herein, refers to an aliphatic groupcontaining at least one triple bond and is intended to include both“unsubstituted alkynyls” and “substituted alkynyls”, the latter of whichrefers to alkynyl moieties having substituents replacing a hydrogen onone or more carbons of the alkynyl group. Such substituents may occur onone or more carbons that are included or not included in one or moretriple bonds. Moreover, such substituents include all those contemplatedfor alkyl groups, as discussed above, except where stability isprohibitive. For example, substitution of alkynyl groups by one or morealkyl carbocyclyl, aryl, heterocyclyl, or heteroaryl groups iscontemplated.

The term “amide”, as used herein, refers to a group

wherein each R¹⁰ independently represent a hydrogen or hydrocarbylgroup, or two R¹⁰ are taken together with the N atom to which they areattached complete a heterocycle having from 4 to 8 atoms in the ringstructure.

The terms “amine” and “amino” are art-recognized and refer to bothunsubstituted and substituted amines and salts thereof, e.g., a moietythat can be represented by

wherein each R¹⁰ independently represents a hydrogen or a hydrocarbylgroup, or two R¹⁰ are taken together with the N atom to which they areattached complete a heterocycle having from 4 to 8 atoms in the ringstructure.

The term “aminoalkyl”, as used herein, refers to an alkyl groupsubstituted with an amino group.

The term “aralkyl”, as used herein, refers to an alkyl group substitutedwith an aryl group.

The term “aryl” as used herein include substituted or unsubstitutedsingle-ring aromatic groups in which each atom of the ring is carbon.Preferably the ring is a 5- to 7-membered ring, more preferably6-membered ring. The term “aryl” also includes polycyclic ring systemshaving two or more cyclic rings in which two or more carbons are commonto two adjoining rings wherein at least one of the rings is aromatic,e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls,cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Aryl groupsinclude benzene, naphthalene, phenanthrene, phenol, aniline, and thelike.

The term “carbamate” is art-recognized and refers to a group

wherein R⁹ and R¹⁰ independently represent hydrogen or a hydrocarbylgroup, such as an alkyl group, or R⁹ and R¹⁰ taken together with theintervening atom(s) complete a heterocycle having from 4 to 8 atoms inthe ring structure.

The terms “carbocycle” and “carbocyclic” as used herein, refers to asaturated or unsaturated ring in which each atom of the ring is carbon.The term carbocycle includes both aromatic carbocycles and non-aromaticcarbocycles. Non-aromatic carbocycles include both cycloalkane rings, inwhich all carbon atoms are saturated, and cycloalkene rings, whichcontain at least one double bond. “Carbocycle” includes 5-7 memberedmonocyclic and 8-12 membered bicyclic rings. Each ring of a bicycliccarbocycle may be selected from saturated, unsaturated and aromaticrings. Carbocycle includes bicyclic molecules in which one, two or threeor more atoms are shared between the two rings. The term “fusedcarbocycle” refers to a bicyclic carbocycle in which each of the ringsshares two adjacent atoms with the other ring. Each ring of a fusedcarbocycle may be selected from saturated, unsaturated and aromaticrings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, maybe fused to a saturated or unsaturated ring, e.g., cyclohexane,cyclopentane, or cyclohexene. Any combination of saturated, unsaturatedand aromatic bicyclic rings, as valence permits, is included in thedefinition of carbocyclic. Exemplary “carbocycles” include cyclopentane,cyclohexane, bicyclo[2.2]heptane, 1,5-cyctooctadiene,1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene andadamantane. Exemplary fused carbocycles include decalin, naphthalene,1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane,4,5,6,7-tetrahydro-1H-indene and bicyclo[4.1.0]hept-3-ene. “Carbocycles”may be substituted at any one or more positions capable of bearing ahydrogen atom.

A “cycloalkyl” group is a cyclic hydrocarbon which is completelysaturated. “Cycloalkyl” includes monocyclic and bicyclic rings.Typically, a monocyclic cycloalkyl group has from 3 to about 10 carbonatoms, more typically 3 to 8 carbon atoms unless otherwise defined. Thesecond ring of a bicyclic cycloalkyl may be selected from saturated,unsaturated and aromatic rings. Cycloalkyl includes bicyclic moleculesin which one, two or three or more atoms are shared between the tworings. The term “fused cycloalkyl” refers to a bicyclic cycloalkyl inwhich each of the rings shares two adjacent atoms with the other ring.The second ring of a fused bicyclic cycloalkyl may be selected fromsaturated, unsaturated and aromatic rings. A “cycloalkenyl” group is acyclic hydrocarbon containing one or more double bonds.

The term “carbocyclylalkyl”, as used herein, refers to an alkyl groupsubstituted with a carbocycle group.

The term “carbonate” is art-recognized and refers to a group —OCO₂—R¹⁰,wherein R¹⁰ represents a hydrocarbyl group.

The term “carboxy”, as used herein, refers to a group represented by theformula —CO₂H.

The term “ester”, as used herein, refers to a group —C(O)OR¹⁰ whereinR¹⁰ represents a hydrocarbyl group.

The term “ether”, as used herein, refers to a hydrocarbyl group linkedthrough an oxygen to another hydrocarbyl group. Accordingly, an ethersubstituent of a hydrocarbyl group may be hydrocarbyl-O—. Ethers may beeither symmetrical or unsymmetrical. Examples of ethers include, but arenot limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethersinclude “alkoxyalkyl” groups, which may be represented by the generalformula alkyl-O-alkyl.

The terms “halo” and “halogen” as used herein means halogen and includeschloro, fluoro, bromo, and iodo.

The terms “hetaralkyl” and “heteroaralkyl”, as used herein, refers to analkyl group substituted with a hetaryl group.

The term “heteroalkyl”, as used herein, refers to a saturated orunsaturated chain of carbon atoms and at least one heteroatom, whereinno two heteroatoms are adjacent. In analogy with alkyl groups,heteroalkyl groups with two open valences are sometimes referred to asheteroalkylene groups. Preferably, the heteroatoms in heteroalkyl groupsare selected from O and N.

The terms “heteroaryl” and “hetaryl” include substituted orunsubstituted aromatic single ring structures, preferably 5- to7-membered rings, more preferably 5- to 6-membered rings, whose ringstructures include at least one heteroatom, preferably one to fourheteroatoms, more preferably one or two heteroatoms. The terms“heteroaryl” and “hetaryl” also include polycyclic ring systems havingtwo or more cyclic rings in which two or more carbons are common to twoadjoining rings wherein at least one of the rings is heteroaromatic,e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls,cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heteroarylgroups include, for example, pyrrole, furan, thiophene, imidazole,oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, andpyrimidine, and the like.

The term “heteroatom” as used herein means an atom of any element otherthan carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, andsulfur.

The terms “heterocyclyl”, “heterocycle”, and “heterocyclic” refer tosubstituted or unsubstituted non-aromatic ring structures, preferably 3-to 10-membered rings, more preferably 3- to 7-membered rings, whose ringstructures include at least one heteroatom, preferably one to fourheteroatoms, more preferably one or two heteroatoms. The terms“heterocyclyl” and “heterocyclic” also include polycyclic ring systemshaving two or more cyclic rings in which two or more carbons are commonto two adjoining rings wherein at least one of the rings isheterocyclic, e.g., the other cyclic rings can be cycloalkyls,cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls.Heterocyclyl groups include, for example, piperidine, piperazine,pyrrolidine, morpholine, lactones, lactams, and the like.

The term “heterocyclylalkyl”, as used herein, refers to an alkyl groupsubstituted with a heterocycle group.

The term “hydrocarbyl”, as used herein, refers to a group that is bondedthrough a carbon atom that does not have a ═O or ═S substituent, andtypically has at least one carbon-hydrogen bond and a primarily carbonbackbone, but may optionally include heteroatoms. Thus, groups likemethyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered to behydrocarbyl for the purposes of this application, but substituents suchas acetyl (which has a ═O substituent on the linking carbon) and ethoxy(which is linked through oxygen, not carbon) are not. Hydrocarbyl groupsinclude, but are not limited to aryl, heteroaryl, carbocycle,heterocyclyl, alkyl, alkenyl, alkynyl, and combinations thereof.

The term “hydroxyalkyl”, as used herein, refers to an alkyl groupsubstituted with a hydroxy group.

The term “lower” when used in conjunction with a chemical moiety, suchas, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant toinclude groups where there are ten or fewer non-hydrogen atoms in thesubstituent, preferably six or fewer. A “lower alkyl”, for example,refers to an alkyl group that contains ten or fewer carbon atoms,preferably six or fewer. In certain embodiments, acyl, acyloxy, alkyl,alkenyl, alkynyl, or alkoxy substituents defined herein are respectivelylower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, orlower alkoxy, whether they appear alone or in combination with othersubstituents, such as in the recitations hydroxyalkyl and aralkyl (inwhich case, for example, the atoms within the aryl group are not countedwhen counting the carbon atoms in the alkyl substituent).

The terms “polycyclyl”, “polycycle”, and “polycyclic” refer to two ormore rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls,heteroaryls, and/or heterocyclyls) in which two or more atoms are commonto two adjoining rings, e.g., the rings are “fused rings”. Each of therings of the polycycle can be substituted or unsubstituted. In certainembodiments, each ring of the polycycle contains from 3 to 10 atoms inthe ring, preferably from 5 to 7. When a polycyclic substituent isattached through an aryl or heteroaryl ring, that substituent may bereferred to herein as an aryl or heteroaryl group, while if thepolycyclic substituent is attached through a cycloalkyl or heterocyclylgroup, that substituent may be referred to herein as a cycloalkyl orheterocyclyl group. By way of example, a1,2,3,4-tetrahydronaphthalen-1-yl group would be a cycloalkyl group,while a 1,2,3,4-tetrahydronaphthalen-5-yl group would be an aryl group.

The term “silyl” refers to a silicon moiety with three hydrocarbylmoieties attached thereto.

The term “substituted” refers to moieties having substituents replacinga hydrogen on one or more carbons or heteroatoms of the moiety. It willbe understood that “substitution” or “substituted with” includes theimplicit proviso that such substitution is in accordance with permittedvalence of the substituted atom and the substituent, and that thesubstitution results in a stable compound, e.g., which does notspontaneously undergo transformation such as by rearrangement,cyclization, elimination, etc. As used herein, the term “substituted” iscontemplated to include all permissible substituents of organiccompounds.

In a broad aspect, the permissible substituents include acyclic andcyclic, branched and unbranched, carbocyclic and heterocyclic, aromaticand non-aromatic substituents of organic compounds. The permissiblesubstituents can be one or more and the same or different forappropriate organic compounds. For purposes of this invention, theheteroatoms such as nitrogen may have hydrogen substituents and/or anypermissible substituents of organic compounds described herein whichsatisfy the valences of the heteroatoms. Substituents can include anysubstituents described herein, for example, a halogen, a hydroxyl, acarbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl),a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate),an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, anamino, an amido, an amidine, an imine, a cyano, a nitro, an azido, asulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, asulfonamide, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic orheteroaromatic moiety, it will be understood by those skilled in the artthat substituents can themselves be substituted, if appropriate. Unlessspecifically stated as “unsubstituted,” references to chemical moietiesherein are understood to include substituted variants. For example,reference to an “aryl” group or moiety implicitly includes bothsubstituted and unsubstituted variants.

The term “sulfate” is art-recognized and refers to the group —OSO₃H, ora pharmaceutically acceptable salt thereof.

The term “sulfonamide” is art-recognized and refers to the grouprepresented by the general formulae

wherein R⁹ and R¹⁰ independently represents hydrogen or hydrocarbyl,such as alkyl, or R⁹ and R¹⁰ taken together with the intervening atom(s)complete a heterocycle having from 4 to 8 atoms in the ring structure.

The term “sulfoxide” is art-recognized and refers to the group—S(O)—R¹⁰, wherein R¹⁰ represents a hydrocarbyl.

The term “sulfonate” is art-recognized and refers to the group SO₃H, ora pharmaceutically acceptable salt thereof.

The term “sulfone” is art-recognized and refers to the group —S(O)₂—R¹⁰,wherein R¹⁰ represents a hydrocarbyl.

The term “thioalkyl”, as used herein, refers to an alkyl groupsubstituted with a thiol group.

The term “thioester”, as used herein, refers to a group —C(O)SR¹⁰ or—SC(O)R¹⁰ wherein R¹⁰ represents a hydrocarbyl.

The term “thioether”, as used herein, is equivalent to an ether, whereinthe oxygen is replaced with a sulfur.

The term “urea” is art-recognized and may be represented by the generalformula

wherein R⁹ and R¹⁰ independently represent hydrogen or a hydrocarbyl,such as alkyl, or either occurrence of R⁹ taken together with R¹⁰ andthe intervening atom(s) complete a heterocycle having from 4 to 8 atomsin the ring structure.

“Protecting group” refers to a group of atoms that, when attached to areactive functional group in a molecule, mask, reduce or prevent thereactivity of the functional group. Typically, a protecting group may beselectively removed as desired during the course of a synthesis.Examples of protecting groups can be found in Greene and Wuts,Protective Groups in Organic Chemistry, 3rd Ed., 1999, John Wiley &Sons, NY and Harrison et al., Compendium of Synthetic Organic Methods,Vols. 1-8, 1971-1996, John Wiley & Sons, NY. Representative nitrogenprotecting groups include, but are not limited to, formyl, acetyl,trifluoroacetyl, benzyl, benzyloxycarbonyl (“CBZ”), tert-butoxycarbonyl(“Boc”), trimethylsilyl (“TMS”), 2-trimethylsilyl-ethanesulfonyl(“TES”), trityl and substituted trityl groups, allyloxycarbonyl,9-fluorenylmethyloxycarbonyl (“FMOC”), nitro-veratryloxycarbonyl(“NVOC”) and the like. Representative hydroxylprotecting groups include,but are not limited to, those where the hydroxyl group is eitheracylated (esterified) or alkylated such as benzyl and trityl ethers, aswell as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers(e.g., TMS or TIPS groups), glycol ethers, such as ethylene glycol andpropylene glycol derivatives and allyl ethers.

As used herein, a therapeutic that “prevents” a disorder or conditionrefers to a compound that, in a statistical sample, reduces theoccurrence of the disorder or condition in the treated sample relativeto an untreated control sample, or delays the onset or reduces theseverity of one or more symptoms of the disorder or condition relativeto the untreated control sample.

The term “treating” includes prophylactic and/or therapeutic treatments.The term “prophylactic or therapeutic” treatment is art-recognized andincludes administration to the host of one or more of the subjectcompositions. If it is administered prior to clinical manifestation ofthe unwanted condition (e.g., disease or other unwanted state of thehost animal) then the treatment is prophylactic (i.e., it protects thehost against developing the unwanted condition), whereas if it isadministered after manifestation of the unwanted condition, thetreatment is therapeutic (i.e., it is intended to diminish, ameliorate,or stabilize the existing unwanted condition or side effects thereof).

The term “prodrug” is intended to encompass compounds which, underphysiologic conditions, are converted into the therapeutically activeagents of the present invention (e.g., a compound of formula I or II). Acommon method for making a prodrug is to include one or more selectedmoieties which are hydrolyzed under physiologic conditions to reveal thedesired molecule. In other embodiments, the prodrug is converted by anenzymatic activity of the host animal. For example, esters or carbonates(e.g., esters or carbonates of alcohols or carboxylic acids) arepreferred prodrugs of the present invention. In certain embodiments,some or all of the compounds of formula I or II in a formulationrepresented above can be replaced with the corresponding suitableprodrug, e.g., wherein a hydroxyl in the parent compound is presented asan ester or a carbonate or carboxylic acid present in the parentcompound is presented as an ester.

Pharmaceutical Compositions

The compositions and methods of the present invention may be utilized totreat an individual in need thereof. In certain embodiments, theindividual is a mammal such as a human, or a non-human mammal. Whenadministered to an animal, such as a human, the composition or thecompound is preferably administered as a pharmaceutical compositioncomprising, for example, a compound of the invention and apharmaceutically acceptable carrier. Pharmaceutically acceptablecarriers are well known in the art and include, for example, aqueoussolutions such as water or physiologically buffered saline or othersolvents or vehicles such as glycols, glycerol, oils such as olive oil,or injectable organic esters. In a preferred embodiment, when suchpharmaceutical compositions are for human administration, particularlyfor invasive routes of administration (i.e., routes, such as injectionor implantation, that circumvent transport or diffusion through anepithelial barrier), the aqueous solution is pyrogen-free, orsubstantially pyrogen-free. The excipients can be chosen, for example,to effect delayed release of an agent or to selectively target one ormore cells, tissues or organs. The pharmaceutical composition can be indosage unit form such as tablet, capsule (including sprinkle capsule andgelatin capsule), granule, lyophile for reconstitution, powder,solution, syrup, suppository, injection or the like. The composition canalso be present in a transdermal delivery system, e.g., a skin patch.

A pharmaceutical composition disclosed herein may comprise a therapeuticcompound in an amount sufficient to allow customary administration to anindividual. In certain embodiments, a pharmaceutical compositiondisclosed herein may comprise, e.g., at least 5 mg, at least 10 mg, atleast 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35mg, at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg, atleast 60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least 80mg, at least 85 mg, at least 90 mg, at least 95 mg, or at least 100 mgof a therapeutic compound. In certain embodiments, a pharmaceuticalcomposition disclosed herein may comprise, e.g., at least 5 mg, at least10 mg, at least 20 mg, at least 25 mg, at least 50 mg, at least 75 mg,at least 100 mg, at least 200 mg, at least 300 mg, at least 400 mg, atleast 500 mg, at least 600 mg, at least 700 mg, at least 800 mg, atleast 900 mg, at least 1,000 mg, at least 1,100 mg, at least 1,200 mg,at least 1,300 mg, at least 1,400 mg, or at least 1,500 mg of atherapeutic compound. In yet other aspects of this embodiment, apharmaceutical composition disclosed herein may comprise in the rangeof, e.g., about 5 mg to about 100 mg, about 10 mg to about 100 mg, about50 mg to about 150 mg, about 100 mg to about 250 mg, about 150 mg toabout 350 mg, about 250 mg to about 500 mg, about 350 mg to about 600mg, about 500 mg to about 750 mg, about 600 mg to about 900 mg, about750 mg to about 1,000 mg, about 850 mg to about 1,200 mg, or about 1,000mg to about 1,500 mg. In still certain embodiments, a pharmaceuticalcomposition disclosed herein may comprise in the range of, e.g., about10 mg to about 250 mg, about 10 mg to about 500 mg, about 10 mg to about750 mg, about 10 mg to about 1,000 mg, about 10 mg to about 1,500 mg,about 50 mg to about 250 mg, about 50 mg to about 500 mg, about 50 mg toabout 750 mg, about 50 mg to about 1,000 mg, about 50 mg to about 1,500mg, about 100 mg to about 250 mg, about 100 mg to about 500 mg, about100 mg to about 750 mg, about 100 mg to about 1,000 mg, about 100 mg toabout 1,500 mg, about 200 mg to about 500 mg, about 200 mg to about 750mg, about 200 mg to about 1,000 mg, about 200 mg to about 1,500 mg,about 5 mg to about 1,500 mg, about 5 mg to about 1,000 mg, or about 5mg to about 250 mg.

A pharmaceutical composition disclosed herein may comprise a solvent,emulsion or other diluent in an amount sufficient to dissolve atherapeutic compound disclosed herein. In certain embodiments, apharmaceutical composition disclosed herein may comprise a solvent,emulsion or a diluent in an amount of e.g., less than about 90% (v/v),less than about 80% (v/v), less than about 70% (v/v), less than about65% (v/v), less than about 60% (v/v), less than about 55% (v/v), lessthan about 50% (v/v), less than about 45% (v/v), less than about 40%(v/v), less than about 35% (v/v), less than about 30% (v/v), less thanabout 25% (v/v), less than about 20% (v/v), less than about 15% (v/v),less than about 10% (v/v), less than about 5% (v/v), or less than about1% (v/v). In certain embodiments, a pharmaceutical composition disclosedherein may comprise a solvent, emulstion or other diluent in an amountin a range of, e.g., about 1% (v/v) to 90% (v/v), about 1% (v/v) to 70%(v/v), about 1% (v/v) to 60% (v/v), about 1% (v/v) to 50% (v/v), about 1% (v/v) to 40% (v/v), about 1% (v/v) to 30% (v/v), about 1% (v/v) to 20%(v/v), about 1% (v/v) to 10% (v/v), about 2% (v/v) to 50% (v/v), about2% (v/v) to 40% (v/v), about 2% (v/v) to 30% (v/v), about 2% (v/v) to20% (v/v), about 2% (v/v) to 10% (v/v), about 4% (v/v) to 50% (v/v),about 4% (v/v) to 40% (v/v), about 4% (v/v) to 30% (v/v), about 4% (v/v)to 20% (v/v), about 4% (v/v) to 10% (v/v), about 6% (v/v) to 50% (v/v),about 6% (v/v) to 40% (v/v), about 6% (v/v) to 30% (v/v), about 6% (v/v)to 20% (v/v), about 6% (v/v) to 10% (v/v), about 8% (v/v) to 50% (v/v),about 8% (v/v) to 40% (v/v), about 8% (v/v) to 30% (v/v), about 8% (v/v)to 20% (v/v), about 8% (v/v) to 15% (v/v), or about 8% (v/v) to 12%(v/v).

The final concentration of a therapeutic compound disclosed herein in apharmaceutical composition disclosed herein may be of any suitableconcentration. In certain embodiments, the final concentration of atherapeutic compound in a pharmaceutical composition may be atherapeutically effective amount. In certain embodiments, the finalconcentration of a therapeutic compound in a pharmaceutical compositionmay be, e.g., at least 0.00001 mg/mL, at least 0.0001 mg/mL, at least0.001 mg/mL, at least 0.01 mg/mL, at least 0.1 mg/mL, at least 1 mg/mL,at least 10 mg/mL, at least 25 mg/mL, at least 50 mg/mL, at least 100mg/mL, at least 200 mg/mL, at least 500 mg/mL, at least 700 mg/mL, atleast 1,000 mg/mL, or at least 1,200 mg/mL. In certain embodiments, theconcentration of a therapeutic compound disclosed herein in the solutionmay be, e.g., at most 1,000 mg/mL, at most 1,100 mg/mL, at most 1,200mg/mL, at most 1,300 mg/mL, at most 1,400 mg/mL, at most 1,500 mg/mL, atmost 2,000 mg/mL, at most 2,000 mg/mL, or at most 3,000 mg/mL. Incertain embodiments, the final concentration of a therapeutic compoundin a pharmaceutical composition may be in a range of, e.g., about0.00001 mg/mL to about 3,000 mg/mL, about 0.0001 mg/mL to about 3,000mg/mL, about 0.01 mg/mL to about 3,000 mg/mL, about 0.1 mg/mL to about3,000 mg/mL, about 1 mg/mL to about 3,000 mg/mL, about 250 mg/mL toabout 3,000 mg/mL, about 500 mg/mL to about 3,000 mg/mL, about 750 mg/mLto about 3,000 mg/mL, about 1,000 mg/mL to about 3,000 mg/mL, about 100mg/mL to about 2,000 mg/mL, about 250 mg/mL to about 2,000 mg/mL, about500 mg/mL to about 2,000 mg/mL, about 750 mg/mL to about 2,000 mg/mL,about 1,000 mg/mL to about 2,000 mg/mL, about 100 mg/mL to about 1,500mg/mL, about 250 mg/mL to about 1,500 mg/mL, about 500 mg/mL to about1,500 mg/mL, about 750 mg/mL to about 1,500 mg/mL, about 1,000 mg/ml, toabout 1,500 mg/mL, about 100 mg/mL to about 1,200 mg/mL, about 250 mg/mLto about 1,200 mg/mL, about 500 mg/mL to about 1,200 mg/mL, about 750mg/mL to about 1,200 mg/mL, about 1,000 mg/mL to about 1,200 mg/mL,about 100 mg/mL to about 1,000 mg/mL, about 250 mg/mL to about 1,000mg/mL, about 500 mg/mL to about 1,000mg/mL, about 750 mg/mL to about1,000 mg/mL, about 100 mg/mL to about 750 mg/mL, about 250 mg/mL toabout 750 mg/mL, about 500 mg/mL to about 750 mg/mL, about 100 mg/mL toabout 500 mg/mL, about 250 mg/mL to about 500 mg/mL, about 0.00001 mg/mLto about 0.0001 mg/mL, about 0.00001 mg/mL to about 0.001 mg/mL, about0.00001 mg/mL to about 0.01 mg/mL, about 0.00001 mg/mL to about 0.1mg/mL, about 0.00001 mg/mL to about 1 mg/mL, about 0.001 mg/mL to about0.01 mg/mL, about 0.001 mg/mL to about 0.1 mg/mL, about 0.001 mg/mL toabout 1 mg/mL, about 0.001 mg/mL to about 10 mg/mL, or about 0.001 mg/mLto about 100 mg/mL.

In certain embodiments, a therapeutically effective amount of atherapeutic compound disclosed herein generally is in the range of about0.001 mg/kg/day to about 100 mg/kg/day. In certain embodiments, aneffective amount of a therapeutic compound disclosed herein may be,e.g., at least 0.001 mg/kg/day, at least 0.01 mg/kg/day, at least 0.1mg/kg/day, at least 1.0 mg/kg/day, at least 5.0 mg/kg/day, at least 10mg/kg/day, at least 15 mg/kg/day, at least 20 mg/kg/day, at least 25mg/kg/day, at least 30 mg/kg/day, at least 35 mg/kg/day, at least 40mg/kg/day, at least 45 mg/kg/day, or at least 50 mg/kg/day. In certainembodiments, an effective amount of a therapeutic compound disclosedherein may be in the range of, e.g., about 0.001 mg/kg/day to about 10mg/kg/day, about 0.001 mg/kg/day to about 15 mg/kg/day, about 0.001mg/kg/day to about 20 mg/kg/day, about 0.001 mg/kg/day to about 25mg/kg/day, about 0.001 mg/kg/day to about 30 mg/kg/day, about 0.001mg/kg/day to about 35 mg/kg/day, about 0.001 mg/kg/day to about 40mg/kg/day, about 0.001 mg/kg/day to about 45 mg/kg/day, about 0.001mg/kg/day to about 50 mg/kg/day, about 0.001 mg/kg/day to about 75mg/kg/day, or about 0.001 mg/kg/day to about 100 mg/kg/day. In yet otheraspects of this embodiment, an effective amount of a therapeuticcompound disclosed herein may be in the range of, e.g., about 0.01mg/kg/day to about 10 mg/kg/day, about 0.01 mg/kg/day to about 15mg/kg/day, about 0.01 mg/kg/day to about 20 mg/kg/day, about 0.01mg/kg/day to about 25 mg/kg/day, about 0.01 mg/kg/day to about 30mg/kg/day, about 0.01 mg/kg/day to about 35 mg/kg/day, about 0.01mg/kg/day to about 40 mg/kg/day, about 0.01 mg/kg/day to about 45mg/kg/day, about 0.01 mg/kg/day to about 50 mg/kg/day, about 0.01mg/kg/day to about 75 mg/kg/day, or about 0.01 mg/kg/day to about 100mg/kg/day. In certain embodiments, an effective amount of a therapeuticcompound disclosed herein may be in the range of, e.g., about 0.1mg/kg/day to about 10 mg/kg/day, about 0.1 mg/kg/day to about 15mg/kg/day, about 0.1 mg/kg/day to about 20 mg/kg/day, about 0.1mg/kg/day to about 25 mg/kg/day, about 0.1 mg/kg/day to about 30mg/kg/day, about 0.1 mg/kg/day to about 35 mg/kg/day, about 0.1mg/kg/day to about 40 mg/kg/day, about 0.1 mg/kg/day to about 45mg/kg/day, about 0.1 mg/kg/day to about 50 mg/kg/day, about 0.1mg/kg/day to about 75 mg/kg/day, or about 0.1 mg/kg/day to about 100mg/kg/day.

In other aspects of this embodiment, an effective amount of atherapeutic compound disclosed herein may be in the range of, e.g.,about 1 mg/kg/day to about 10 mg/kg/day, about 1 mg/kg/day to about 15mg/kg/day, about 1 mg/kg/day to about 20 mg/kg/day, about 1 mg/kg/day toabout 25 mg/kg/day, about 1 mg/kg/day to about 30 mg/kg/day, about 1mg/kg/day to about 35 mg/kg/day, about 1 mg/kg/day to about 40mg/kg/day, about 1 mg/kg/day to about 45 mg/kg/day, about 1 mg/kg/day toabout 50 mg/kg/day, about 1 mg/kg/day to about 75 mg/kg/day, or about 1mg/kg/day to about 100 mg/kg/day. In certain embodiments, an effectiveamount of a therapeutic compound disclosed herein may be in the rangeof, e.g., about 5 mg/kg/day to about 10 mg/kg/day, about 5 mg/kg/day toabout 15 mg/kg/day, about 5 mg/kg/day to about 20 mg/kg/day, about 5mg/kg/day to about 25 mg/kg/day, about 5 mg/kg/day to about 30mg/kg/day, about 5 mg/kg/day to about 35 mg/kg/day, about 5 mg/kg/day toabout 40 mg/kg/day, about 5 mg/kg/day to about 45 mg/kg/day, about 5mg/kg/day to about 50 mg/kg/day, about 5 mg/kg/day to about 75mg/kg/day, or about 5 mg/kg/day to about 100 mg/kg/day.

In liquid and semi-solid formulations, a concentration of a therapeuticcompound disclosed herein typically may be between about 50 mg/mL toabout 1,000 mg/mL. In certain embodiments, a therapeutically effectiveamount of a therapeutic disclosed herein may be from, e.g., about 50mg/ml, to about 100 mg/mL, about 50 mg/mL to about 200 mg/mL, about 50mg/mL to about 300 mg/mL, about 50 mg/mL to about 400 mg/mL, about 50mg/mL to about 500 mg/mL, about 50 mg/mL to about 600 mg/mL, about 50mg/mL to about 700 mg/mL, about 50 mg/mL to about 800 mg/mL, about 50mg/mL to about 900 mg/mL, about 50 mg/mL to about 1,000 mg/mL, about 100mg/mL to about 200 mg/mL, about 100 mg/mL to about 300 mg/mL, about 100mg/mL to about 400 mg/mL, about 100 mg/mL to about 500 mg/mL, about 100mg/mL to about 600 mg/mL, about 100 mg/mL to about 700 mg/mL, about 100mg/mL to about 800 mg/mL, about 100 mg/mL to about 900 mg/mL, about 100mg/mL to about 1,000 mg/mL, about 200 mg/mL to about 300 mg/mL, about200 mg/mL to about 400 mg/mL, about 200 mg/mL to about 500 mg/mL, about200 mg/mL to about 600 mg/mL, about 200 mg/mL to about 700 mg/mL, about200 mg/mL to about 800 mg/mL, about 200 mg/mL to about 900 mg/mL, about200 mg/mL to about 1,000 mg/mL, about 300 mg/mL to about 400 mg/mL,about 300 mg/mL to about 500 mg/mL, about 300 mg/mL to about 600 mg/mL,about 300 mg/mL to about 700 mg/mL, about 300 mg/mL to about 800 mg/mL,about 300 mg/mL to about 900 mg/mL, about 300 mg/mL to about 1,000mg/mL, about 400 mg/mL to about 500 mg/mL, about 400 mg/mL to about 600mg/mL, about 400 mg/mL to about 700 mg/mL, about 400 mg/mL to about 800mg/mL, about 400 mg/mL to about 900 mg/mL, about 400 mg/mL to about1,000 mg/mL, about 500 mg/mL to about 600 mg/mL, about 500 mg/mL toabout 700 mg/mL, about 500 mg/mL to about 800 mg/mL, about 500 mg/mL toabout 900 mg/mL, about 500 mg/mL to about 1,000 mg/mL, about 600 mg/mLto about 700 mg/mL, about 600 mg/mL to about 800 mg/mL, about 600 mg/mLto about 900 mg/mL, or about 600 mg/mL to about 1,000 mg/mL.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the ability of certain compounds of the invention tomitigate radiation induced apoptosis in vitro relative to controlvalues.

FIG. 2 shows the mitigation against whole body radiation lethality invivo by compound 1 (FIG. 2A), compound 2 (FIG. 2B), compound 3 (FIG.2C), compound 4 (FIG. 2D), compound 5 (FIG. 2E).

FIG. 3 shows the dose response experiments for compounds 2 (5355512) and3 (5346360) in C3H male mice and C57B1/6 female mice.

FIG. 4 shows the dose response experiments for compounds 2 (5355512) and3 (5346360) when increased to 25 mg/kg (FIG. 4A and 4B) and if dosing ofcompound 2 (5355512) was given after 24 hours (D1), 48 hours (D2), and72 hours (D3) (FIG. 4C).

FIG. 5 shows the dose response when compounds 2 (5355512) and 3(5346360) are given orally by gavage.

FIG. 6 shows the Probit analysis of the effect of varying the radiationdose with compounds 2 (5355512), 3 (5346360), and 5 (5116319).

FIG. 7 shows the mitigation against whole body radiation lethality invivo by analogs of Compound 2 (5355512).

FIG. 8 shows the reduction in radiation induced apoptosis with theadministration of certain compounds of the invention.

FIG. 9 shows flow cytometric assay results demonstrating the ability ofactive compounds to mitigate radiation-induced apoptosis relative toinactive compounds.

FIG. 10 shows that compounds 2 (5355512) and 3 (5346360) inhibit TNF-αproduction when added to stimulated peritoneal macrophages 1 hour afterLPS measured by ELISA (FIG. 10A) and other compounds of the inventionthat were able to inhibit TNF-α production (FIG. 10B)

FIG. 11 shows that compounds 2 (5355512) and 3 (5346360) inhibitexpression of TNF-α mRNA, and other cytokines, when added to bonemarrow-derived macrophages 1 hour after LPS measured by RT-PCR.

FIG. 12 shows how compound 2 (5355512) induces peritoneal macrophages toproduce TNF-α in response to lipopolysaccharide (LPS).

FIG. 13 shows that compound 2 (5355512) increases the total endogenousCFU in spleens (FIG. 13A) and average endogenous CPU per spleen (FIG.13B) at 10 days after various doses of whole body irradiation (WBI).

FIG. 14 shows the results of the in vivo assay with compound 2 (5355512)in C57B1/6 mice who were irradiated with 16 and 18 Gy abdominally.

FIG. 15 shows the mitigation of radiation pneumonitis and fibrosis afterlocal thoracic irradiation in C3H mice with pneumonitis (FIG. 15A) andC57B1/6 mice with fibrosis (FIG. 15B).

FIG. 16 shows the effects of the certain compounds of the invention onthe growth of tumor lung colonies in vivo and on radiation response.

DETAILED DESCRIPTION

Compounds shown in Table 1 mitigate the effects of ionizing radiation,inhibit inflammation, and are useful for treating cancer and otherhyperproliferative disorders. Pharmaceutical formulations usingcompounds shown in Table 1 have potential to improve the outcome ofradiation exposure, and therefore they may be useful in the cancerradiotherapy, as well as in the situation of a radiological emergency.

Radiomitigation

Without wishing to be bound by theory, the compounds shown in Table 1may protect against deleterious effects of ionizing radiation bypromoting repair of DNA damage caused by exposure to radiation. Thecompounds also inhibit inflammation as well as protect bone marrow andother organs from radiation damage. As disclosed herein, after in vitroscreening, the chosen compounds were applied in mice 24 hours after TBIand provided greatly elevated animals' survival level, compared to acontrol group. They also mitigated lethal normal gut and lung radiationdamage, as well as having anti-tumor activity with and withouttherapeutic doses of ionizing radiation.

Embodiments of the invention include pharmaceutical formulations havinga compound shown in Table 1, or pharmaceutically acceptable saltthereof, and at least one pharmaceutically acceptable carrier ordiluent.

Embodiments of the invention include methods of mitigating the effect ofionizing radiation on a cell, organ, tissue, or organism by contactingthe cell, organ, tissue, or organism with at least one compound shown inTable 1.

As used herein, “mitigating” means reducing the negative effects causedby exposure to ionizing radiation, relative to a cell, organ, tissue, ororganism exposed to the same level of radiation for the same amount oftime, but untreated.

In some embodiments, contacting the cell, organ, tissue, or organismwith a compound in Table 1 may comprise administering a therapeuticallyeffective amount of the compound to a subject.

As used herein, a “therapeutically effective amount” is an amountsufficient to mitigate the effects of the ionizing radiation.

The subject may be any organism that has been exposed to ionizingradiation, or which may be exposed to ionizing radiation. In oneembodiment, the invention provides a method wherein the subject is ahuman, rat, mouse, cat, dog, horse, sheep, cow, monkey, avian, oramphibian. In another embodiment, the cell is in vivo or in vitro.Typical subjects to which compounds of the invention may be administeredwill be mammals, particularly primates, especially humans. Forveterinary applications, a wide variety of subjects will be suitable, e.g. livestock such as cattle, sheep, goats, cows, swine and the like;poultry such as chickens, ducks, geese, turkeys, and the like; anddomesticated animals particularly pets such as dogs and cats. Fordiagnostic or research applications, a wide variety of mammals will besuitable subjects including rodents (e.g. mice, rats, hamsters),rabbits, primates, and swine such as inbred pigs and the like.Additionally, for in vitro applications, such as in vitro diagnostic andresearch applications, body fluids and cell samples of the abovesubjects will be suitable for use such as mammalian, particularlyprimate such as human, blood, urine or tissue samples, or blood urine ortissue samples of the animals mentioned for veterinary applications.

The cell, organ, tissue, or organism may be contacted with a compoundshown in Table 1 before, during, or after exposure to ionizingradiation. In some embodiments, the compound may be administeredprophylactically, i.e. before exposure to ionizing radiation, forexample, prior to cancer radiation therapy or X-ray. In someembodiments, the compound may be administered during exposure, or uponrepeated exposure to ionizing radiation. In some embodiments, thecompound may be administered after exposure to ionizing radiation, orafter the initiation of exposure to radiation.

When administering to an organism, the compound may be administered byany suitable means. In some embodiments, the compounds or formulationsare administered orally. In some embodiments, the compounds orformulations are administered by injection, e.g. subcutaneous,parenteral, or intravenous, injections. In some embodiments the compoundmay be administered in combination with other potential mitigators orwith other toxic agents such as chemotherapeutic drugs.

Ionizing radiation may refer to radiation with a photon energy greaterthan 10 eV, according to the U.S. Federal Communications Commission, butfor biological purposes may be considered to be radiation having energygreater than the first ionization potential of oxygen or the ionizationpotential of hydrogen, and may have other meanings according topractitioners.

Inflammatory Diseases

Compounds of the current invention may also be used for the treatment orprevention of inflammation and inflammatory diseases.

Examples of inflammatory conditions, which may be treated or preventedby the administration of a compound of the invention include, but arenot limited to, inflammation of the lungs, joints, connective tissue,eyes, nose, bowel, kidney, liver, skin, central nervous system, vascularsystem and heart. In certain embodiments, inflammatory conditions whichmay be treated by the current invention include inflammation due to theinfiltration of leukocytes or other immune effector cells into affectedtissue. Other relevant examples of inflammatory conditions which may betreated by the present invention include inflammation caused byinfectious agents, including, but not limited to, viruses, bacteriafungi and parasites.

Inflammatory lung conditions include, but are not limited to, asthma,adult respiratory distress syndrome, bronchitis, pulmonary inflammation,pulmonary fibrosis, and cystic fibrosis (which may additionally oralternatively involve the gastro-intestinal tract or other tissue(s)).Inflammatory joint conditions include rheumatoid arthritis, rheumatoidspondylitis, juvenile rheumatoid arthritis, osteoarthritis, goutyarthritis and other arthritic conditions. Eye diseases with aninflammatory component include, but are not limited to, uveitis(including iritis), conjunctivitis, scleritis, keratoconjunctivitissicca, and retinal diseases, including, but not limited to, diabeticretinopathy, retinopathy of prematurity, retinitis pigmentosa, and dryand wet age-related macular degeneration. Inflammatory bowel conditionsinclude Crohn's disease, ulcerative colitis and distal proctitis.

Inflammatory skin diseases include, but are not limited to, conditionsassociated with cell proliferation, such as psoriasis, eczema anddermatitis, (e.g., eczematous dermatitides, topic and seborrheicdermatitis, allergic or irritant contact dermatitis, eczema craquelee,photoallergic dermatitis, phototoxic dermatitis, phytophotodermatitis,radiation dermatitis, and stasis dermatitis). Other inflammatory skindiseases include, but are not limited to, scleroderma, ulcers anderosions resulting from trauma, burns, bullous disorders, or ischemia ofthe skin or mucous membranes, several forms of ichthyoses, epidermolysisbullosae, hypertrophic scars, keloids, cutaneous changes of intrinsicaging, photoaging, frictional blistering caused by mechanical shearingof the skin and cutaneous atrophy resulting from the topical use ofcorticosteroids. Additional inflammatory skin conditions includeinflammation of mucous membranes, such as cheilitis, chapped lips, nasalirritation, mucositis and vulvovaginitis.

Inflammatory disorders of the endocrine system include, but are notlimited to, autoimmune thyroiditis (Hashimoto's disease), Type Idiabetes, and acute and chronic inflammation of the adrenal cortex.Inflammatory conditions of the cardiovascular system include, but arenot limited to, coronary infarct damage, peripheral vascular disease,myocarditis, vasculitis, revascularization of stenosis, atherosclerosis,and vascular disease associated with Type II diabetes.

Inflammatory condition of the kidney include, but are not limited to,glomerulonephritis, interstitial nephritis, lupus nephritis, nephritissecondary to Wegener's disease, acute renal failure secondary to acutenephritis, Goodpasture's syndrome, post-obstructive syndrome and tubularischemia.

Inflammatory conditions of the liver include, but are not limited to,hepatitis (arising from viral infection, autoimmune responses, drugtreatments, toxins, environmental agents, or as a secondary consequenceof a primary disorder), biliary atresia, primary biliary cirrhosis andprimary sclerosing cholangitis.

Inflammatory conditions of the central nervous system include, but arenot limited to, multiple sclerosis and neurodegenerative diseases suchas Alzheimer's disease, Parkinson's disease, or dementia associated withHIV infection.

Other inflammatory conditions include periodontal disease, tissuenecrosis in chronic inflammation, endotoxin shock, smooth muscleproliferation disorders, graft versus host disease, tissue damagefollowing ischemia reperfusion injury, and tissue rejection followingtransplant surgery.

The present invention further provides a method of treating orpreventing inflammation associated with post-surgical wound healing in apatient comprising administering to said patient a compound of theinvention.

It should be noted that compounds of the current invention may be usedto treat or prevent any disease which has an inflammatory component,such as those diseases cited above. Further, the inflammatory conditionscited above are meant to be exemplary rather than exhaustive.

Those skilled in the art would recognize that additional inflammatoryconditions (e.g., systemic or local immune imbalance or dysfunction dueto an injury, an insult, infection, inherited disorder, or anenvironmental intoxicant or perturbant to the subject's physiology) maybe treated or prevented by compounds of the current invention. Thus, themethods of the current invention may be used to treat or prevent anydisease which has an inflammatory component, including, but not limitedto, those diseases cited above.

The present invention also provides methods for treating or preventingarthritis, inflammatory bowel disease, uveitis, ocular inflammation,asthma, pulmonary inflammation, cystic fibrosis, psoriasis, arterialinflammation, cardiovascular diseases, multiple sclerosis, orneurodegenerative disease by administering an effective amount of acompound of the invention.

The present invention also provides methods for treating ischemia byadministering an effective amount of a compound of the invention. Incertain embodiments, the ischemia is cardiac ischemia, cerebralischemia, bowel ischemia (e.g., ischemic colitis or mesentericischemia), or cutaneous ischemia.

Cancer

Compounds of the current invention may also be used for the treatment ofcancer. The actual symptoms associated with cancer are well known andcan be determined by a person of ordinary skill in the art by takinginto account one or more factors, including, without limitation, thelocation of the cancer, the cause of the cancer, the severity of thecancer, and/or the tissue or organ affected by the cancer. Those ofskill in the art will know the appropriate symptoms or indicatorsassociated with a specific type of cancer and will know how to determineif an individual is a candidate for treatment as disclosed herein.

Exemplary forms of cancer which may be treated by the subject methodsinclude, but are not limited to, leukemia, non-Hodgkin's lymphoma,prostate cancer, bladder cancer, lung cancer (including either smallcell or non-small cell cancer), colon cancer, kidney cancer, livercancer, breast cancer, cervical cancer, endometrial or other uterinecancer, ovarian cancer, skin cancer (e.g., melanoma), testicular cancer,cancer of the penis, cancer of the vagina, cancer of the urethra, gallbladder cancer, esophageal cancer, or pancreatic cancer. Additionalexemplary forms of cancer which may be treated by the subject methodsinclude, but are not limited to, cancer of skeletal or smooth muscle,stomach cancer, cancer of the small intestine, cancer of the salivarygland, anal cancer, rectal cancer, thyroid cancer, parathyroid cancer,pituitary cancer, and nasopharyngeal cancer.

In certain embodiments, a therapeutic compound disclosed herein reducesthe size of a tumor by, e.g., at least 10%, at least 15%, at least 20%,at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 55%, at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90% or at least 95%. Inyet other aspects of this embodiment, a therapeutic compound disclosedherein reduces the size of a tumor from, e.g., about 5% to about 100%,about 10% to about 100%, about 20% to about 100%, about 30% to about100%, about 40% to about 100%, about 50% to about 100%, about 60% toabout 100%, about 70% to about 100%, about 80% to about 100%, about 10%to about 90%, about 20% to about 90%, about 30% to about 90%, about 40%to about 90%, about 50% to about 90%, about 60% to about 90%, about 70%to about 90%, about 10% to about 80%, about 20% to about 80%, about 30%to about 80%, about 40% to about 80%, about 50% to about 80%, or about60% to about 80%, about 10% to about 70%, about 20% to about 70%, about30% to about 70%, about 40% to about 70%, or about 50% to about 70%.

In certain embodiments, a cancer therapeutic disclosed herein is capableof reducing the number of cancer cells in an individual suffering from acancer by, e.g., at least 10%, at least 15%, at least 20%, at least 25%,at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 55%, at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90% or at least 95% as compared to apatient not receiving the same treatment. In other aspects of thisembodiment, a cancer therapeutic is capable of reducing the number ofcancer cells in an individual suffering from a cancer by, e.g., about10% to about 100%, about 20% to about 100%, about 30% to about 100%,about 40% to about 100%, about 50% to about 100%, about 60% to about100%, about 70% to about 100%, about 80% to about 100%, about 10% toabout 90%, about 20% to about 90%, about 30% to about 90%, about 40% toabout 90%, about 50% to about 90%, about 60% to about 90%, about 70% toabout 90%, about 10% to about 80%, about 20% to about 80%, about 30% toabout 80%, about 40% to about 80%, about 50% to about 80%, or about 60%to about 80%, about 10% to about 70%, about 20% to about 70%, about 30%to about 70%, about 40% to about 70%, or about 50% to about 70% ascompared to a patient not receiving the same treatment.

In certain embodiments, a therapeutically effective amount of a cancertherapeutic disclosed herein reduces or maintains a cancer cellpopulation and/or tumor cell size in an individual by, e.g., at least10%, at least 15%, at least 20%, at least 25%, at least 30%, at least35%, at least 40%, at least 45%, at least 50%, at least 55%, at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95% or at least 100%. In other aspects ofthis embodiment, a therapeutically effective amount of a cancertherapeutic disclosed herein reduces or maintains a cancer cellpopulation and/or tumor cell size in an individual by, e.g., at most10%, at most 15%, at most 20%, at most 25%, at most 30%, at most 35%, atmost 40%, at most 45%, at most 50%, at most 55%, at most 60%, at most65%, at most 70%, at most 75%, at most 80%, at most 85%, at most 90%, atmost 95% or at most 100%. In yet other aspects of this embodiment, atherapeutically effective amount of a cancer therapeutic disclosedherein reduces or maintains a cancer cell population and/or tumor cellsize in an individual by, e.g., about 10% to about 100%, about 10% toabout 90%, about 10% to about 80%, about 10% to about 70%, about 10% toabout 60%, about 10% to about 50%, about 10% to about 40%, about 20% toabout 100%, about 20% to about 90%, about 20% to about 80%, about 20% toabout 20%, about 20% to about 60%, about 20% to about 50%, about 20% toabout 40%, about 30% to about 100%, about 30% to about 90%, about 30% toabout 80%, about 30% to about 70%, about 30% to about 60%, or about 30%to about 50%.

Compounds

The compounds herein described may have one or more asymmetric centersor planes. Compounds of the present invention containing anasymmetrically substituted atom may be isolated in optically active orracemic forms. It is well known in the art how to prepare opticallyactive forms, such as by resolution of racemic forms (racemates), byasymmetric synthesis, or by synthesis from optically active startingmaterials. Resolution of the racemates can be accomplished, for example,by conventional methods such as crystallization in the presence of aresolving agent, or chromatography, using, for example a chiral HPLCcolumn. Many geometric isomers of olefins, C═N double bonds, and thelike can also be present in the compounds described herein, and all suchstable isomers are contemplated in the present invention. Cis and transgeometric isomers of the compounds of the present invention aredescribed and may be isolated as a mixture of isomers or as separatedisomeric forms. All chiral (enantiomeric and diastereomeric), andracemic forms, as well as all geometric isomeric forms of a structureare intended, unless the specific stereochemistry or isomeric form isspecifically indicated.

The compounds herein described may have one or more charged atoms. Forexample, the compounds may be zwitterionic, but may be neutral overall.Other embodiments may have one or more charged groups, depending on thepH and other factors. In these embodiments, the compound may beassociated with a suitable counter-ion. It is well known in the art howto prepare salts or exchange counter-ions. Generally, such salts can beprepared by reacting free acid forms of these compounds with astoichiometric amount of the appropriate base (such as Na, Ca, Mg, or Khydroxide, carbonate, bicarbonate, or the like), or by reacting freebase forms of these compounds with a stoichiometric amount of theappropriate acid. Such reactions are typically carried out in water orin an organic solvent, or in a mixture of the two. Counter-ions may bechanged, for example, by ion-exchange techniques such as ion-exchangechromatography. All zwitterions, salts and counter-ions are intended,unless the counter-ion or salt is specifically indicated. In certainembodiments, the salt or counter-ion may be pharmaceutically acceptable,for administration to a subject. Pharmaceutically acceptable salts arediscussed later.

Pharmaceutical Compositions

Embodiments of the invention include pharmaceutical compositions ofcompounds shown in Table 1 and at least one pharmaceutically acceptablecarrier or diluent. As used herein, pharmaceutical compositions includecompositions suitable for administration to a subject or patient. Assuch, compositions do not include chemical reaction solutions orsolutions used for screening assays, as these are not suitable foradministration to a subject or patient. In some embodiments thecompositions may include one or more than one compound from Table 1, oneor more other pharmaceutically active agent, and may further containother suitable substances and excipients, including but not limited tophysiologically acceptable buffering agents, stabilizers (e.g.antioxidants), flavoring agents, agents to effect the solubilization ofthe compound, and the like.

In other embodiments, the composition may be in any suitable form suchas a solution, a suspension, an emulsion, an infusion device, or adelivery device for implantation or it may be presented as a dry powderto be reconstituted with water or another suitable vehicle before use.The composition may include suitable pharmaceutically acceptablecarriers and/or excipients.

In other embodiments, the compositions may comprise an effective amountof a modulator and/or other pharmaceutically active agent in aphysiologically-acceptable carrier. The carrier may take a wide varietyof forms depending on the form of preparation desired for a particularroute of administration. Suitable carriers and their formulation aredescribed, for example, in Remington's Pharmaceutical Sciences by E. W.Martin.

In some embodiments, the compound may be contained in any appropriateamount in any suitable carrier substance, and is generally present in anamount of 1-95% by weight of the total weight of the composition. Thecomposition may be provided in a dosage form that is suitable forparenteral (e.g., subcutaneously, intravenously, intramuscularly, orintraperitoneally) or oral administration route. The pharmaceuticalcompositions may be formulated according to conventional pharmaceuticalpractice (see, e.g., Remington: The Science and Practice of Pharmacy(20th ed.), ed, A. R. Gennaro, Lippincott Williams & Wilkins, 2000 andEncyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C.Boylan, 1988-1999, Marcel Dekker, New York).

In some embodiments, the compositions may be in a form suitable foradministration by sterile injection. In one example, to prepare such acomposition, the composition(s) are dissolved or suspended in aparenterally acceptable liquid vehicle. Among acceptable vehicles andsolvents that may be employed are water, water adjusted to a suitable pHby addition of an appropriate amount of hydrochloric acid, sodiumhydroxide or a suitable buffer, 1,3-butanediol, Ringer's solution, andisotonic sodium chloride solution and dextrose solution. The aqueousformulation may also contain one or more preservatives (e.g., methyl,ethyl or n-propyl p-hydroxybenzoate). For parenteral formulations, thecarrier will usually comprise sterile water, though other ingredients,for example, ingredients that aid solubility or for preservation, may beincluded, injectable solutions may also be prepared in which caseappropriate stabilizing agents may be employed.

Formulations suitable for parenteral administration usually comprise asterile aqueous preparation of the compound, which may be isotonic withthe blood of the recipient (e.g., physiological saline solution). Suchformulations may include suspending agents and thickening agents andliposomes or other microparticulate systems which are designed to targetthe compound to blood components or one or more organs. The formulationsmay be presented in unit-dose or multi-dose form.

Parenteral administration may comprise any suitable form of systemicdelivery or localized delivery. Administration may for example beintravenous, intra-arterial, intrathecal, intramuscular, subcutaneous,intramuscular, intra-abdominal (e.g., intraperitoneal), etc., and may beeffected by infusion pumps (external or implantable) or any othersuitable means appropriate to the desired administration modality.

In some embodiments, the compositions may be in a form suitable for oraladministration. In compositions in oral dosage form, any of the usualpharmaceutical media may be employed. Thus, for liquid oralpreparations, such as, for example, suspensions, elixirs and solutions,suitable carriers and additives include water, glycols, oils, alcohols,flavoring agents, preservatives, coloring agents and the like. For solidoral preparations such as, for example, powders, capsules and tablets,suitable carriers and additives include starches, sugars, diluents,granulating agents, lubricants, binders, disintegrating agents and thelike. If desired, tablets may be sugar coated or enteric coated bystandard techniques.

Compositions suitable for oral administration may be presented asdiscrete units such as capsules, cachets, tablets, or lozenges, eachcontaining a predetermined amount of the active ingredient as a powderor granules. Optionally, a suspension in an aqueous liquor or anon-aqueous liquid may be employed, such as a syrup, an elixir, anemulsion, or a draught. Formulations for oral use include tabletscontaining active ingredient(s) in a mixture with pharmaceuticallyacceptable excipients. Such formulations are known to the skilledartisan. Excipients may be, for example, inert diluents or fillers(e.g., sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose,starches including potato starch, calcium carbonate, sodium chloride,lactose, calcium phosphate, calcium sulfate, or sodium phosphate);granulating and disintegrating agents (e.g., cellulose derivativesincluding microcrystalline cellulose, starches including potato starch,croscarmellose sodium, alginates, or alginic acid); binding agents(e.g., sucrose, glucose, sorbitol, acacia, alginic acid, sodiumalginate, gelatin, starch, pregelatinized starch, microcrystallinecellulose, magnesium aluminum silicate, carboxymethylcellulose sodium,methylcellulose, hydroxypropyl methylcellulose, ethylcellulose,polyvinylpyrrolidone, or polyethylene glycol); and lubricating agents,glidants, and antiadhesives (e.g., magnesium stearate, zinc stearate,stearic acid, silicas, hydrogenated vegetable oils, or talc). Otherpharmaceutically acceptable excipients can be colorants, flavoringagents, plasticizers, humectants, buffering agents, and the like.

A syrup may be made by adding the compound to a concentrated aqueoussolution of a sugar, for example sucrose, to which may also be added anyaccessory ingredient(s). Such accessory ingredient(s) may includeflavorings, suitable preservative, agents to retard crystallization ofthe sugar, and agents to increase the solubility of any otheringredient, such as a polyhydroxy alcohol, for example glycerol orsorbitol.

In some embodiments, the composition may be in a form of nasal or othermucosal spray formulations (e.g. inhalable forms). These formulationscan include purified aqueous solutions of the active compounds withpreservative agents and isotonic agents. Such formulations can beadjusted to a pH and isotonic state compatible with the nasal or othermucous membranes. Alternatively, they can be in the form of finelydivided solid powders suspended in a gas carrier. Such formulations maybe delivered by any suitable means or method, e.g., by nebulizer,atomizer, metered dose inhaler, or the like.

In some embodiments, the composition may be in a form suitable forrectal administration. These formulations may be presented as asuppository with a suitable carrier such as cocoa butter, hydrogenatedfats, or hydrogenated fatty carboxylic acids.

In some embodiments, the composition may be in a form suitable fortransdermal administration. These formulations may be prepared, forexample, by incorporating the active compound in a thixotropic orgelatinous carrier such as a cellulosic medium, e.g., methyl celluloseor hydroxyethyl cellulose, with the resulting formulation then beingpacked in a transdermal device adapted to be secured in dermal contactwith the skin of a wearer.

In addition to the aforementioned ingredients, compositions of theinvention may further include one or more accessory ingredient(s)selected from encapsulants, diluents, buffers, flavoring agents,hinders, disintegrants, surface active agents, thickeners, lubricants,preservatives (including antioxidants), and the like.

in some embodiments, compositions may be formulated for immediaterelease, sustained release, delayed-onset release or any other releaseprofile known to one skilled in the art.

In some embodiments, the pharmaceutical composition may be formulated torelease the active compound substantially immediately uponadministration or at any predetermined time or time period afteradministration. The latter types of compositions are generally known ascontrolled release formulations, which include (i) formulations thatcreate a substantially constant concentration of the drug within thebody over an extended period of time; (ii) formulations that after apredetermined lag time create a substantially constant concentration ofthe drug within the body over an extended period of time; (iii)formulations that sustain action during a predetermined time period bymaintaining a relatively constant, effective level in the body withconcomitant minimization of undesirable side effects associated withfluctuations in the plasma level of the active substance (sawtoothkinetic pattern); (iv) formulations that localize action, by, e.g.,spatial placement of a controlled release composition adjacent to or inthe central nervous system or cerebrospinal fluid; (v) formulations thatallow for convenient dosing, such that doses are administered, forexample, once every one or two weeks; and (vi) formulations that targetthe site of a pathology. For some applications, controlled releaseformulations obviate the need for frequent dosing to sustain activity ata medically advantageous level.

Any of a number of strategies can be pursued in order to obtaincontrolled release in which the rate of release outweighs the rate ofmetabolism of the compound in question. In one example, controlledrelease is obtained by appropriate selection of various formulationparameters and ingredients, including, e.g., various types of controlledrelease compositions and coatings. Thus, the compound is formulated withappropriate excipients into a pharmaceutical composition that, uponadministration, releases the compound in a controlled manner. Examplesinclude single or multiple unit tablet or capsule compositions, oilsolutions, suspensions, emulsions, microcapsules, microspheres,molecular complexes, nanoparticles, patches, and liposomes.

In some embodiments, the composition may comprise a “vectorized” form,such as by encapsulation of the compound in a liposome or otherencapsulate medium, or by fixation of the compound, e.g., by covalentbonding, chelation, or associative coordination, on a suitablebiomolecule, such as those selected from proteins, lipoproteins,glycoproteins, and polysaccharides.

In some embodiments, the composition can be incorporated intomicrospheres, microcapsules, nanoparticles, liposomes, or the like forcontrolled release. Furthermore, the composition may include suspending,solubilizing, stabilizing, pH-adjusting agents, tonicity adjustingagents, and/or dispersing, agents. Alternatively, the compound may beincorporated in biocompatible carriers, implants, or infusion devices.

Materials for use in the preparation of microspheres and/ormicrocapsules are, e.g., biodegradable/bioerodible polymers such aspolygalactin, poly-(isobutyl cyanoacrylate),poly(2-hydroxyethyl-L-glutamine) and, poly(lactic acid). Biocompatiblecarriers that may be used when formulating a controlled releaseparenteral formulation are carbohydrates (e.g., dextrans), proteins(e.g., albumin), lipoproteins, or antibodies. Materials for use inimplants can be non-biodegradable (e.g., polydimethyl siloxane) orbiodegradable (e.g., poly(caprolactone), poly(lactic acid),poly(glycolic acid) or poly(ortho esters) or combinations thereof).

In all embodiments, the compound or other active compounds may bepresent as pharmaceutically acceptable salts or other derivatives, suchas ether derivatives, ester derivatives, acid derivatives, and aqueoussolubility altering derivatives of the active compound. Derivativesinclude all individual enantiomers, diastereomers, racemates, and otherisomers of the compounds. Derivatives also include all polymorphs andsolvates, such as hydrates and those formed with organic solvents, ofthe compounds. Such isomers, polymorphs, and solvates may be prepared bymethods known in the art, such as by regiospecific and/orenantioselective synthesis and resolution.

The ability to prepare salts depends on the acidity or basicity of thecompounds. Suitable salts of the compounds include, but are not limitedto, acid addition salts, such as those made with hydrochloric,hydrobromic, hydroiodic, perchloric, sulfuric, nitric, phosphoric,acetic, propionic, glycolic, lactic pyruvic, malonic, succinic, maleic,fumaric, malic, tartaric, citric, benzoic, carbonic, cinnamic, mandelic,methanesulfonic, ethanesulfonic, hydroxyethanesulfonic,benezenesulfonic, p-toluene sulfonic, cyclohexanesulfamic, salicyclic,p-aminosalicylic, 2-phenoxybenzoic, and 2-acetoxybenzoic acid; saltsmade with saccharin; alkali metal salts, such as sodium and potassiumsalts; alkaline earth metal salts, such as calcium and magnesium salts;and salts formed with organic or inorganic ligands, such as quaternaryammonium salts.

Additional suitable salts include, but are not limited to, acetate,benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate,bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate,citrate, dihydrochloride, edetate, edisylate, estolate, esylate,fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate,hexylresorcinate, hydrabamine, hydrobromide, hydrochloride,hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate,malate, maleate, mandelate, mesylate, methylbromide, methylnitrate,methylsulfate, mucate, napsylate, nitrate, N-methylglucamine ammoniumsalt, oleate, pamoate (embonate), palmitate, pantothenate,phosphate/diphosphate, polygalacturonate, salicylate, stearate, sulfate,subacetate, succinate, tannate, tartrate, teoclate, tosylate,triethiodide and valerate salts of the compounds.

The pharmaceutically acceptable acid addition salts can also exist asvarious solvates, such as with water, methanol, ethanol,dimethylformamide, and the like. Mixtures of such solvates can also beprepared. The source of such solvate can be from the solvent ofcrystallization, inherent in the solvent of preparation orcrystallization, or adventitious to such solvent.

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: (1)water-soluble antioxidants, such as ascorbic acid, cysteinehydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfiteand the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate,butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT),lecithin, propyl gallate, alpha-tocopherol, and the like; and (3)metal-chelating agents, such as citric acid, ethylenediamine tetraaceticacid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Unless the context clearly indicates otherwise, compositions of allembodiments can comprise various pharmaceutically acceptable salts, orother derivatives described above.

The formulation and preparation of such compositions are well known tothose skilled in the art of pharmaceutical formulation. Formulations canbe found in Remington: The Science and Practice of Pharmacy.

The amount of the compound employed in the present invention to be usedvaries according to the degree of the exposure to ionizing radiationencountered, and the stages of any radiation-induced damage. A suitabledosage is that which will result in concentration of the compound (inblood and/or tissues) sufficient to mitigate the damage of the ionizingradiation. The preferred dosage is that amount sufficient to render asubject asymptomatic after exposure to ionizing radiation.

The contents of all cited references (including literature references,issued patents, published patent applications) as cited throughout thisapplication are hereby expressly incorporated by reference. Theinvention and the manner and process of making and using it, aredescribed in such full, clear, concise and exact terms as to enable anyperson skilled in the art to which it pertains, to make and use thesame.

The term “unit dosage form” or “unit” as used herein refers tophysically discrete units suitable as unitary dosages for human andanimal subjects, each unit containing a predetermined quantity of thecompound calculated in an amount sufficient to produce the desiredeffect in association with a pharmaceutically acceptable, diluent,carrier or vehicle. The specifications for the novel unit dosage formsof the present invention depend on the particular compound employed andthe effect to be achieved, and the pharmacodynamics associated with eachcompound in the subject.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions maybe varied so as to obtain an amount of the activeingredient that is effective to achieve the desired therapeutic responsefor a particular patient, composition, and mode of administration,without being toxic to the patient.

The selected dosage level will depend upon a variety of factorsincluding the activity of the particular compound or combination ofcompounds employed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion of theparticular compound(s) being employed, the duration of the treatment.other drugs, compounds and/or materials used in combination with theparticular compound(s) employed, the age, sex, weight, condition,general health and prior medical history of the patient being treated,and like factors well known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readilydetermine and prescribe the therapeutically effective amount of thepharmaceutical composition required. For example, the physician orveterinarian could start doses of the pharmaceutical composition orcompound at levels lower than that required in order to achieve thedesired therapeutic effect and gradually increase the dosage until thedesired effect is achieved. By “therapeutically effective amount” ismeant the concentration of a compound that is sufficient to elicit thedesired therapeutic effect. It is generally understood that theeffective amount of the compound will vary according to the weight, sex,age, and medical history of the subject. Other factors which influencethe effective amount may include, but are not limited to, the severityof the patient's condition, the disorder being treated, the stability ofthe compound, and, if desired, another type of therapeutic agent beingadministered with the compound of the invention. A larger total dose canbe delivered by multiple administrations of the agent. Methods todetermine efficacy and dosage are known to those skilled in the art(Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13ed., 1814-1882, herein incorporated by reference).

In general, a suitable daily dose of an active compound used in thecompositions and methods of the invention will be that amount of thecompound that is the lowest dose effective to produce a therapeuticeffect. Such an effective dose will generally depend upon the factorsdescribed above.

Dosing can be single dosage or cumulative (serial dosing), and can bereadily determined by one skilled in the art. For instance, treatmentmay comprise a one-time administration of an effective dose of apharmaceutical composition disclosed herein. Alternatively, treatmentmay comprise multiple administrations of an effective dose of apharmaceutical composition earned out over a range of time periods, suchas, e.g., once daily, twice daily, thrice daily, once every few days, oronce weekly. The timing of administration can vary from individual toindividual, depending upon such factors as the severity of anindividual's symptoms. For example, an effective dose of apharmaceutical composition disclosed herein can be administered to anindividual once daily for an indefinite period of time, or until theindividual no longer requires therapy. A person of ordinary skill in theart will recognize that the condition of the individual can be monitoredthroughout the course of treatment and that the effective amount of apharmaceutical composition disclosed herein that is administered can beadjusted accordingly.

If desired, the effective daily dose of the active compound may beadministered as one, two, three, four, five, six or more sub-dosesadministered separately at appropriate intervals throughout the day,optionally, in unit dosage forms. In certain embodiments of the presentinvention, the active compound may be administered two or three timesdaily. In preferred embodiments, the active compound will beadministered once daily.

In certain embodiments, the period of administration of a therapeuticcompound is for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks,12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10months, 11 months, 12 months, or more. In certain embodiments, atreatment regimen may comprise a period during which administration isstopped for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks,12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10months, 11 months, 12 months, or more.

The patient receiving this treatment is any animal in need, includingprimates, in particular humans, and other mammals such as equines,cattle, swine and sheep; and poultry and pets in general.

In other embodiments, the compound of Formula I or II may be providedwith the one or more additional therapeutic agents in a kit, e.g., asseparate pharmaceutical formulations capable of being used together in aconjoint therapy as discussed herein, either together in a singlecontainer or in separate containers. In certain such embodiments, thekit may further include instructions for the conjoint administration ofthe pharmaceutical formulations, e.g., for treating or preventing any ofthe conditions discussed above.

Such combination products may employ compounds of this invention, orpharmaceutically acceptable salts thereof within the dosage rangedescribed hereinbefore and the other pharmaceutically-active agentwithin its approved dosage range.

The invention now being generally described, it will be more readilyunderstood by reference to the following examples which are includedmerely for purposes of illustration of certain aspects and embodimentsof the present invention, and are not intended to limit the invention.

EXAMPLES Example 1

The structures of 12 4-nitrophenylsulfonylpiperazines (NPSPs) or4-nitrophenylsulfonamides (NPSs) are shown. See Table 1

TABLE 1 The structures of 12 NPSPs or NPSs of the invention Com- Com-pound pound ID Compound Structure Compound Name 1 5348300

1-(4-fluorophenyl)- 4-[(4-nitrobenzene) sulfonyl]piperazine 2 5355512

1-[(4- nitrobenzene)sulfonyl]- 4-phenylpiperazine 3 5346360

1-(3-chlorophenyl)- 4-(4- nitrobenzene)sulfonyl) piperazine 4 5347486

1-[(4-nitrobenzene) Sulfonyl]-4- [(2E)-3-phenylprop-2-en-1-yl]piperazine 5 5116319

N,N-diethyl-4- nitrophenyl-1- sulfonamide 6 5475972

3-((4- nitrophenyl)sulfonyl)- 2-phenyltetrahydro- thiophene 7 5344400

1-methyl-4-((4- nitrophenyl)sulfo- nyl)piperazine 8 6561181

1-((4- nitrophenyl)sulfonyl)- 4-(3- phenoxybenzyl)piper- azine 9 5243457

4-benzyl-1-((4- nitrophenyl)sulfo- nyl)piperidine 10 6571802

1-(4- (methylthio)benzyl)- 4-((4-nitrophen- yl)sulfonyl)piper- azine 11AST5814142

3-cyano,7-methyl- 2-(4-((4-nitrophen- yl)sulfonyl)piper-azin-1-yl)quinoline 12 AST6538836

3-cyano,7,8- dimethyl-2-(4-((4- nitrophenyl)sulfonyl)- 1,4-diazepan-1-yl)quinoline

Example 2

The ability of compounds from Example 1 to mitigate radiation-inducedapoptosis in vitro relative to control values (100%) is shown in FIG. 1.Briefly, 100,000 small-molecules were screened at 30 μM finalconcentration in 1% dimethylsulfoxide (DMSO) using the viability of amurine lymphocyte line as a readout (Mitigation cell viability %). Tenthousand Til-1 murine lymphocyte cells were dispensed into each well of384-well plates using a Multidrop 384 (Thermo Scientific, Waltham,Mass.) The Til-1 cells were irradiated at the dose of 2Gy. After 1 hour,the small-molecules were added. Twenty-four hours after the radiation,cell viability was assayed by luminesce-based measurement of ATPproduction (ATPlite, Perkin-Elmer, Waltham, Mass.) with a SpectraMax M5microplate reader (Molecular Devices, Sunnyvale, Calif.). The Z′ factorfor the assay was >0.5. See Zhang, J. H., Chung, T. D., Oldenburg, K. R.“A Simple Statistical Parameter for Use in Evaluation and Validation ofHigh Throughput Screening Assays,” J Biomol Screen 4:67, 1999. Apositive readout was ≦130 % of the control irradiated value. Compounds1-7 showed activity, including a 4-nitrophenyl sulfonamide derivative(Compound 5). This suggests a minimum moiety for efficacy. Thepiperazine group afforded additional efficacy beyond the commonsulfonamide core suggesting that both moieties contribute although theywere not in themselves sufficient for full activity. See FIG. 1.

Example 3

The top 5 performers from Example 1 were tested in vivo for theirability to mitigate against lethality from whole body irradiation. SeeTable 2.

Briefly, Animal Whole Body Irradiation (WBI) Assays were conducted withC3Hf/Kam and C57B16/J mice. The mice were bred and maintained in astrict gnotobiotic environment in the American Association of LaboratoryAnimal Care-Accredited Animal Facilities of Department of RadiationOncology, UCLA. The Animal Care and Use Committee approved allexperiments, which were performed in accordance with all local andnational guidelines for the care and use of animals. Mice, 9-12 weeksold, were given WBI using a Gamma cell 40 irradiator (Cs137 source:Atomic Energy of Canada, Ltd.) at a dose rate of 67 Gy/min. Mice weremonitored for at least 30 days and defined criteria for humaneeuthanasia was used as an endpoint.

Compounds were typically dissolved in 15 μL DMSO and suspended in 1 mLof 1% Cremphor EL in water for administration in 0.2 mL volumes. Thisamount of Cremophor did not significantly alter the response to WBI. Allmice, including controls, received the same diluent as the experimentalgroups. The chosen compounds were administered to the mice 5 times at 24h intervals, starting 24 hours after WBI at LD 70/30 (7.725 Gy for theC3H strain) doses. Similar data were obtained with a single dose and at48 hours after radiation exposure in both mouse strains. Bothsubcutaneous and oral routes were effective. The compounds providedgreatly elevated animals survival level as compared to control group.

All 5 compounds were effective but Compound 2 (5355512) and Compound 3(5346360) were effective at 5 mg/kg injected subcutaneously whereas theothers required a greater quantity of the tested compound (e.g., 75mg/kg). Indeed, Compound 2 and Compound 3 gave 100% survivors at day 30at the lowest dosage of 5 mg/kg which was superior to the higher dosesof 40 mg/kg and 75 mg/kg. See FIG. 2 A-E.

TABLE 2 Compound Solubility Compound ID In Vitro (mol/L × 10⁻⁶) 15348300 188% 26.6 2 5355512 182% 66.1 3 5346360 174% 6.5 4 5347486 163%20.4 5 5116319 161% 512.9

Example 4

Dose response experiments were executed in a manner similar to the WBIAssay described in Example 3. These experiments indicated that 5 mg/kgwas more, or as, effective, than 1, 2, 10, 40, or 75 mg/kg for bothCompound 2 (5355512) and Compound 3 (5346360) in C3H male mice. See FIG.3. Both Compound 2 (5355512) and Compound 3 (5346360) were effective inC57B1/6 female mice in this case mitigating against LD70/30 doses of8.509 Gy due to the relative radioresistance of this strain relative toC3Hs. A single dose of 5 mg/kg given 24 hours after WBI was effective(See FIGS. 4A and 4B) and increasing the dose to 25 mg/kg did notimprove efficacy. Some activity was retained if a single dose was given48, but not 72, hours after WBI (See FIG. 4C). When given orally bygavage, Compound 2 (5355512) in 5 daily doses starting at 24 hourspost-WBI was effective at 5 mg/kg in both C3H and C57B1/6 strains, butnot at 25 mg/kg. (See FIG. 5) Interestingly, Compound 3 (5346360) givenby the same gavage schedule was inactive in both strains at either dose.

Example 5

Probit analyses of the effect of varying the radiation dose withCompound 2 (5355512) (5 mg/kg), and Compound 3 (5346360) (5 mg/kg), or 5(5116319) (75 mg/kg) given subcutaneously for 5 days starting 24 hoursafter WBI, is shown in FIG. 6, with 95% confidence limits shown for LD50values. The dose-response curves are steep with the dose modifyingfactors varying with the level of effect and drug, but were in the range1.04-1.09. Such factors have to be considered against the level of thecontrol and the reason for lethality. In this case, hematologicalinsufficiency and not infection is the cause of death and as a resultthe radioresistance of the C3H strain is relatively high.

Example 6

Compounds P1, P2, and P3, analogs of Compound 2 (5355512), weresynthesized according to standard protocols. See Table 3. The simplestNPSP (P1) was effective indicating that this was the optimum “core”structure. Interestingly, P1 is a natural breakdown product identifiedin vivo by mass spectrometry. WBI irradiation assays were conductedsimilar to those described in Example 3 except that compounds P1, P2,and P3 are soluble in water. The analogs were injected subcutaneously 5times over 5 days at 5 mg/kg. Compound P2 elicited 100% survival after30 days. See FIG. 7.

TABLE 3 Analogs of 5355512 Compound ID Structure 5355512

Compound P1

Compound P2

Compound P3

Example 7

The ability of active compounds to mitigate radiation-induced apoptosiswas confirmed relative to inactive compounds in a flow cytometric assaythat employed annexin V for early stage apoptosis and propidium iodidefor late stage apoptosis (BD BioSciences). (See FIGS. 8 and 9).

Example 8

NPSPs were tested for their anti-inflammatory ability using murinemacrophages. Compound 2 (5355512) showed anti-inflammatory activitybeing able to inhibit lipopolysaccharide (LPS)-induced production oftumor necrosis factor and other inflammatory cytokines. Briefly,peritoneal macrophages (PMs) were induced by intraperitoneal, injectionof 150 μg M1S416 (Innate Immunotherapeutics, NZ) and harvested on day 4by peritoneal wash out with PBS. Culture supernatants were harvested at24 hrs and cytokines tested by ELISA (BD Biosciences, SD). Compound 2(5355512) and Compound 3 (5346360) inhibit TNF-α production when addedto stimulated peritoneal macrophages 1 hr after LPS measured by ELISA.See FIG. 10A. Additionally, all NPSPs and compound 5 (5116319) were ableto inhibit TNF-α production. See FIG. 10B.

Additionally, bone marrow-derived macrophages (BMDMs) were derived by 7days

culture of marrow ceils in medium containing 10% FBS and CSF-Iconditioned medium. The serum concentration was reduced to 2% FBS 16 hbefore stimulation with LPS for 30 min, treatment with drug andincubation for another 3.5 h (4 h total with LPS). Total cellular RNAwas isolated by trizol and cDNA synthesized using iScript from BioRad.Gene expression was measured by qPCR and analyzed using the standardcurve method, normalized to L32, Compound 2 (5355512) and Compound 3(5346360) inhibit expression of TNF-α mRNA, and other cytokines, whenadded to bone marrow-derived macrophages 1 hr after LPS measured byRT-PCR. See FIG. 11.

In vivo, compound 2 (5355512) showed anti-inflammatory activityreflected by its ability to generate myeloid suppressor cells (data notshown).

To determine if compound 2 (5355512), injected subcutaneously wouldaffect the activation status of induced peritoneal, macrophages,compound 2 (5355512) was given subcutaneously to control or WBI (5 Gy)and peritoneal macrophages harvested at day 4 as before. Cells fromthese groups of mice were incubated for 24 h with LPS or LPS then 512(See FIG. 12). Surprisingly, treatment with Compound 2 (5355512) in vivo“primed” induced peritoneal cells to produce TNF-α in response to LPS toalmost the same extent as WBI alone. However, this “priming” effect wasless when WBI and Compound 2 (5355512) were combined in vivo. Also, inkeeping with the data in FIGS. 10 and 11, the addition of Compound 2(5355512) in vitro blunted TNF-α production.

Example 9

Compound 2 (5355512) also increased the number of endogenous colonyforming units in the spleens of mice that had received LD70/30 radiationdoses which is a measure of ability to generate hematopoietic stem cellsand myeloid suppressors. The compounds increased survival when given 5times at 24 hour intervals, starting 24 hours after abdominal or lungirradiation. For example, Compound 2 (5355512) increased the totalendogenous CFU in spleens and average endogenous CFU per spleen at 10days after whole body irradiation (WBI) at varying doses (e.g. 6.5 Gy7.0 Gy, and 7.5 Gy). (See FIG. 13).

Example 10

Further in vivo experiments similar to Example 3 were conducted except alocalized radiation was used to demonstrate mitigation of abdominalradiation with compound 2 (5355512) at increased radiation doses. SeeFIG. 14 Additionally, in vivo experiments demonstrated mitigation ofradiation pneumonitis and fibrosis after local thoracic irradiation.Addition of compound 2 (5355512) supported survival of C3H mice and C57mice to 100 days post thoracic radiation (14 and 18 Gy). (See FIG. 15)

Example 11

The Lewis lung (LLC) model of artificial metastasis was used to test theeffects of the drug on the growth of tumor lung colonies in vivo and onradiation response. This was chosen because it is easy to see any drugeffects on growth promotion and radioprotection in multiple tumorssimultaneously. Specifically, C57B1/6 mice and C3H mice were injectedintravenously with 5×10⁴ tumor cells. Subcutaneous injections ofcompound 2 (5355512) were started on day 4 when the tumors were alreadyestablished in the lung. A daily dose of 20 mg/kg for 5 days was used tobias the experiment in favor of any tumor growth promotion. Localthoracic irradiation (LTI) was started on day 5 with 4 Gy daily dosesfor 3 days. This is higher than conventional 2 Gy to compensate for therapid growth of murine tumors, but is still well within the higherranges used clinically in hypofractionated exposures. The colonies werecounted on day 14. FIG. 16 shows that compound 2 (5355512) significantly(P<0.05) decreased the number of lung tumor colonies assessed on day 14by 20%. The colonies were also smaller in size. LTI alone decreased thenumber of colonies by 40%. This was not further decreased by thesuboptimal drug treatment. Thus, compounds given to mice bearing LewisLung tumors decreased the number of tumor colonies that grew with andwithout lung irradiation demonstrating that the compounds did notprotect tumors and indicating that they may be useful in patientsreceiving radiation therapy for cancer, protecting normal tissue fromdamage while exerting anti-tumor action.

As described herein, all embodiments or subcombinations may be used incombination with all other embodiments or subcombinations, unlessmutually exclusive.

The embodiments illustrated and discussed in this specification areintended only to teach those skilled in the art the best way known tothe inventors to make and use the invention. Nothing in thisspecification should be considered as limiting the scope of the presentinvention. All examples presented are representative and non-limiting.The above-described embodiments of the invention may be modified orvaried, without departing from the invention, as appreciated by thoseskilled in the art in light of the above teachings. It is therefore tobe understood that, within the scope of the claims and theirequivalents, the invention may be practiced otherwise than asspecifically described.

INCORPORATION BY REFERENCE

All publications and patents mentioned herein are hereby incorporated byreference in their entirety as if each individual publication or patentwas specifically and individually indicated to be incorporated byreference. In case of conflict, the present application, including anydefinitions herein, will control.

EQUIVALENTS

While specific embodiments of the subject invention have been discussed,the above specification is illustrative and not restrictive. Manyvariations of the invention will become apparent to those skilled in theart upon review of this specification and the claims below. The fullscope of the invention should be determined by reference to the claims,along with their full scope of equivalents, and the specification, alongwith such variations.

1. A compound having the structure of Formula I:

wherein: A⁵ is a secondary or tertiary amino substituent, and A⁶ is asubstituted or unsubstituted aryl or heteroaryl group.
 2. A compound ofclaim 1, wherein the aryl or heteroaryl group bears at least onesubstituent including a nitro substitutent.
 3. The compound of claim 2,wherein the nitro substituent is disposed at a position on A⁶ distal tothe sulfonyl.
 4. The compound of any preceding claim 1, wherein A⁵ is aheterocyclic amine.
 5. The compound of claim 1, wherein A⁵ is NR¹⁵R¹⁶,wherein each of R¹⁵ and R¹⁶, independently, is H, alkyl, alkenyl,alkynyl, aryl, heteroaryl, aralkyl, or heteroaralkyl, provided that R¹⁵and R¹⁶ are not both H.
 6. A compound of claim 1, having the structureof Formula II:

wherein: X is N or —C(H)—; and Y¹and Y² are each independently loweralkyl or Y¹ and Y² taken together with X form a heterocyclyl ring. 7.The compound of claim 6, wherein Y¹ and Y² are each ethyl.
 8. Thecompound of claim 6, wherein Y¹ and Y² taken together with X form apiperazine ring.
 9. The compound of claim 6, wherein Y¹ and Y² takentogether with X form:

wherein X is N; G is selected from N or —C(H)—; Z is absent or selectedfrom substituted or unsubstituted alkyl, heteroalkyl, alkenyl, oralkynyl; R⁴ is absent or selected from substituted or unsubstituted aryland heteroaryl; and R⁵ and R⁶ are each independently absent or loweralkyl.
 10. The compound of claim 9, wherein G is N and R⁴ is selectedfrom phenyl, 4-fluorophenyl and 3-chlorophenyl.
 11. The compound ofclaim 9, wherein Z is absent.
 12. The compound of claim 9, wherein Z isprop-2-en-1-yl and R⁴ is phenyl.
 13. The compound of claim 6, wherein Y¹and Y² taken together with X form:

wherein X is —C(H)—, and R⁴ is absent or selected from substituted orunsubstituted aryl and heteroaryl.
 14. A pharmaceutical compositioncomprising a compound of claim 1 and a pharmaceutically acceptableexcipient or solvent.
 15. A method of mitigating an effect of ionizingradiation on a cell, organ, tissue, or organism, comprising contactingthe cell, organ, tissue, or organism with a compound of claim
 1. 16. Themethod of claim 15, wherein the compound contacts the cell before,during, or after exposure to ionizing radiation.
 17. A method oftreating inflammation in an organism, comprising administering to theorganism a compound of claim
 1. 18. A method of treating cancer in anorganism, comprising administering to the organism a compound ofclaim
 1. 19. The method of claim 18, wherein the organism is a mammal.20. The compound of claim 3, wherein A⁶ is optionally substituted4-nitrophenyl.